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Resolving the molecular architecture of the photoreceptor active zone by MINFLUX nanoscopy

View ORCID ProfileChad P. Grabner, Isabelle Jansen, Jakob Neef, Tobias Weiss, Roman Schmidt, Dietmar Riedel, Christian A. Wurm, Tobias Moser
doi: https://doi.org/10.1101/2021.05.28.446138
Chad P. Grabner
1Institute for Auditory Neuroscience, University Medical Center Göttingen, 37075 Göttingen, Germany
2Synaptic Nanophysiology Group, Max Planck Institute for Biophysical Chemistry Göttingen, Germany
3Cluster of Excellence “Multiscale Bioimaging: from molecular machines to networks of excitable cells”, University of Göttingen, 37075 Göttingen, Germany
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  • ORCID record for Chad P. Grabner
Isabelle Jansen
4Abberior Instruments, Hans-Adolf-Krebs-Weg 1, 37077 Göttingen, Germany
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Jakob Neef
1Institute for Auditory Neuroscience, University Medical Center Göttingen, 37075 Göttingen, Germany
2Synaptic Nanophysiology Group, Max Planck Institute for Biophysical Chemistry Göttingen, Germany
3Cluster of Excellence “Multiscale Bioimaging: from molecular machines to networks of excitable cells”, University of Göttingen, 37075 Göttingen, Germany
5Collaborative Research Center 1286, University of Göttingen, Göttingen, Germany
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Tobias Weiss
4Abberior Instruments, Hans-Adolf-Krebs-Weg 1, 37077 Göttingen, Germany
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Roman Schmidt
4Abberior Instruments, Hans-Adolf-Krebs-Weg 1, 37077 Göttingen, Germany
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Dietmar Riedel
6Laboratory of Electron Microscopy, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany
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Christian A. Wurm
4Abberior Instruments, Hans-Adolf-Krebs-Weg 1, 37077 Göttingen, Germany
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  • For correspondence: tmoser@gwdg.de c.wurm@abberior-instruments.com
Tobias Moser
1Institute for Auditory Neuroscience, University Medical Center Göttingen, 37075 Göttingen, Germany
2Synaptic Nanophysiology Group, Max Planck Institute for Biophysical Chemistry Göttingen, Germany
3Cluster of Excellence “Multiscale Bioimaging: from molecular machines to networks of excitable cells”, University of Göttingen, 37075 Göttingen, Germany
5Collaborative Research Center 1286, University of Göttingen, Göttingen, Germany
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  • For correspondence: tmoser@gwdg.de c.wurm@abberior-instruments.com
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Abstract

Cells assemble macromolecular complexes into scaffoldings that serve as substrates for catalytic processes. Years of molecular neurobiology indicate that neurotransmission depends on such optimization strategies, yet the molecular topography of the presynaptic Active Zone (AZ) where transmitter is released upon synaptic vesicle (SV) fusion remains to be visualized. Therefore, we implemented MINFLUX optical nanoscopy to resolve the AZ of rod photoreceptors. To facilitate MINFLUX nanoscopy of the AZ, we developed and verified an immobilization technique, we name Heat Assisted Rapid Dehydration (HARD). Here fresh retinal slices are directly stamped onto glass coverslips yielding a single layer of rod AZs. These AZs exhibited excellent labeling efficiency and minimal signal redundancy in the Z-direction. Our data indicate that the SV release site is a molecular complex of bassoon-Rab3-binding molecule 2 (RIM2)-ubMunc13-2-CAST. The complexes are serially duplicated longitudinally, and reflected in register along the axis of symmetry of the synaptic ribbon.

One sentence summary Structural motifs formed by active zone proteins at the photoreceptor synapse.

Competing Interest Statement

Abberior Instruments develops and manufactures super-resolution fluorescence microscopes, including Confocal, STED and MINFLUX systems.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted May 30, 2021.
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Resolving the molecular architecture of the photoreceptor active zone by MINFLUX nanoscopy
Chad P. Grabner, Isabelle Jansen, Jakob Neef, Tobias Weiss, Roman Schmidt, Dietmar Riedel, Christian A. Wurm, Tobias Moser
bioRxiv 2021.05.28.446138; doi: https://doi.org/10.1101/2021.05.28.446138
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Resolving the molecular architecture of the photoreceptor active zone by MINFLUX nanoscopy
Chad P. Grabner, Isabelle Jansen, Jakob Neef, Tobias Weiss, Roman Schmidt, Dietmar Riedel, Christian A. Wurm, Tobias Moser
bioRxiv 2021.05.28.446138; doi: https://doi.org/10.1101/2021.05.28.446138

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